Pancreatic cancer (PC) has the number one fatality rate of all cancers, and is the fourth leading cause of cancer-related deaths in North America, with an overall five-year survival rate less than 5%. Therefore, there is an urgent need to identify novel molecular targets in PC that could guide the choice of effective therapies. The novel concepts in this proposal are that a zinc transporter, ZIP4, regulates PC cell growth, tumor progression, and survival, which suggests a new and important role for ZIP4. We have also found that ZIP4 is overexpressed in majority of PC specimens and PC cell lines to varying degrees. Forced overexpression of ZIP4 increases PC cell proliferation and tumor growth. Conversely, silencing of ZIP4 by shRNA inhibits PC growth and increases the survival rate in a mouse model, suggesting that ZIP4 is a potential therapeutic target. Our preliminary data also demonstrate that microRNA-224 (miR-224) is downregulated in ZIP4 over-expressing cells and xenografts, and is upregulated when ZIP4 is silenced. A potential target of miR-224, apoptosis inhibitor 5 (API-5), is found to be upregulated in ZIP4 overexpressing PC cells and downregulated in ZIP4 silenced PC cells, suggesting a new mechanism of ZIP4-mediated PC growth. We hypothesize that the aberrant expression of ZIP4 plays a critical role in PC cell growth and tumor progression through the miR-224/API-5 pathway, and ZIP4 may be a novel therapeutic target for PC. We propose to determine whether the expression of ZIP4 a) varies in a K-Ras transgenic mouse model and correlates with tumor progression, b) correlates with zinc levels in a K-Ras transgenic mouse model. We will also determine whether overexpression of ZIP4 a) downregulates the transcription of miR-224, and b) affects PC cell apoptosis through the miR-224/API-5 pathway. Finally, we will determine whether a) 3 cycles of liposome/ZIP4 shRNA treatment, and b) combinational therapy of liposome/ZIP4 shRNA plus gemcitabine is effective on PC in a mouse model. The novel findings in this R01 proposal are that the dietary zinc transporter ZIP4 is over-expressed in majority of patients with PC and regulates PC growth and survival. The proposed studies will help us to understand the correlation of ZIP4 and PC progression by using molecular approaches.